Nanoparticle Coupled Antioxidants for Respiratory Illness in Veterans

描述(由申请人提供)： 在被派往伊拉克或阿富汗的退伍军人中，超过三分之二的人患有呼吸系统疾病。呼吸道疾病是持久自由行动(OEF)和伊拉克自由行动(OIF)退伍军人中最常见的医疗疾病之一，这是一个一贯的发现，影响到军事人员，无论年龄、军衔、性别或现役/国民警卫队身份。近23%的OEF/OIF退伍军人认为单位效能受到呼吸系统疾病的负面影响1。众所周知，即使是轻微的上呼吸道感染也会对军事行动产生重大影响，2有人对部署到伊拉克的人员中呼吸道感染和过敏的频率可能增加表示关切。我们的退伍军人经历了越来越多的环境暴露，这可能会导致这些炎症性呼吸道疾病，包括烟草烟雾和柴油废气。近一半的OEF/OIF退伍军人在部署期间开始或重新开始吸烟。此外，由于柴油发动机在军用车辆和装备中的广泛使用，暴露在柴油废气颗粒物(DEP)中。烟草烟雾和DEP都是已知的空气传播毒素，由于自由基会引起广泛的氧化性炎症，并可能导致过敏性鼻炎的加重3。自由基的快速降解导致活性氧物种(ROS)4-6。代谢ROS的抗氧化酶因此成为潜在的治疗剂。这些抗氧化剂包括超氧化物歧化酶(SOD)和过氧化氢酶(CAT)，前者将毒性最大的ROS之一的超氧化物转化为活性较低的过氧化氢，后者将过氧化氢转化为水和氧气。抗氧化剂治疗使用的一个问题是确保局部使用的抗氧化剂与呼吸道粘膜接触的时间足够长，才能有效。正常的粘液纤毛清除可在15-20分钟内将外用药物从呼吸道中移除，此时保持粘膜接触数小时至数天更为可取。我们建议使用可生物降解的聚乳酸纳米颗粒(NPs)实现对呼吸道上皮细胞中抗氧化剂SOD和CAT(抗氧化剂)的靶向传递和保留。我们将同时用SOD和/或CAT和一种针对粘蛋白1(MUC1)的靶向抗体(Ab)包被NPs，该抗体结合到呼吸道上皮细胞的管腔表面。我们的初步研究表明，HSNECs在调节对环境刺激的炎症反应中起着关键作用，以抗氧化剂为靶点的HSNECs可以有利地影响随后的APC和T细胞反应。我们的中心假设是，OEF/OIF退伍军人经历的烟雾和DEP暴露影响了HSNEC-APC通过ROS的通讯，这种炎症反应可以通过使用针对呼吸道上皮的抗氧化剂纳米颗粒来改善。这一假说将通过以下特定目标来验证：特定目标1：确定抗体-抗氧化剂-纳米粒的结合特性和上皮毒性。用抗MUC1Ab将超氧化物歧化酶和过氧化氢酶与聚乳酸纳米粒同时共价结合。我们将制备不同比例的超氧化物歧化酶、过氧化氢酶和抗MUC1三种不同比例的神经细胞，并用体外培养的HSNEC评价不同比例的超氧化物歧化酶、过氧化氢酶和抗MUC1对其结合特性、功能和毒性的影响。具体目的2：确定抗氧化剂纳米粒体外抑制烟雾和DEP诱导的炎症反应的效果。我们将使用目标1中确定的优化的抗MUC1-SOD/CAT-NPs来确定预防烟雾和DEP在体外诱导的HSNECs和APC炎症反应的有效性和能力，作为免疫反应的初始步骤。具体目的3：确定抗氧化剂纳米粒对吸烟和DEP加重的鼻窦炎的体内疗效。我们将使用一种因吸烟或DEP暴露而加重的变应性真菌性鼻-鼻窦炎小鼠模型来确定抗MUC1-SOD/CAT-NPs的疗效。 公共卫生相关性： 叙述(与退伍军人健康相关)呼吸系统疾病影响着部署在伊拉克或阿富汗的三分之二以上的退伍军人，是持久自由行动(OEF)和伊拉克自由行动(OIF)退伍军人中最常见的医疗条件之一。来自军用车辆的烟草烟雾和柴油废气颗粒中的颗粒物的独特暴露可能是这种异常高发病率的原因之一，并导致呼吸道的氧化性炎症。我们的战略目标是开发一种局部用药，这种药物可以用作治疗措施，以对抗呼吸道上皮的自由基促炎作用，即使在移除这些环境暴露后，这种作用也可以持续下去。我们的中心假设是，与呼吸道粘膜上表达的粘蛋白1蛋白的抗体以及治疗性抗氧化剂、超氧化物和过氧化氢酶结合的纳米颗粒与上皮细胞的接触增加，可以防止继发于烟雾和DEP暴露的损害。

DESCRIPTION (provided by applicant): Respiratory illnesses affect over two-thirds of Veterans deployed to Iraq or Afghanistan. The fact that respiratory tract disorders are among the most common medical conditions in Operation Enduring Freedom (OEF) and Operation Iraqi Freedom (OIF) Veterans has been a consistent finding that affects military personnel regardless of age, rank, gender or active duty/National Guard status. Nearly 23% of OEF/OIF Veterans felt unit effectiveness was negatively affected by respiratory illnesses1. Even minor upper respiratory tract infections are known to have a significant impact on military operations2 and concern has been raised about a possible increase in frequency of respiratory tract infections and allergy among personnel deployed to Iraq1. Our Veterans experience an increased number of environmental exposures that likely contribute to these inflammatory respiratory conditions, to include tobacco smoke and diesel exhaust. Nearly half of OEF/OIF Veterans started or re-started smoking during deployment. Additionally, exposure to diesel exhaust particles (DEP) occurs from the extensive use of diesel engines in military vehicles and equipment. Both tobacco smoke and DEP are known airborne toxins that cause extensive oxidative inflammation due to free radicals and can lead to exacerbation of allergic rhinitis3. Quick degradation of free radicals results in reactive oxygen species (ROS)4-6. Antioxidant enzymes that metabolize ROS, thus serve as potential therapeutic agents. These antioxidants include superoxide dismutase (SOD), which converts superoxide, one of the most toxic ROS, to much less reactive peroxide, and catalase (CAT), which converts peroxide into water and oxygen. One problem with therapeutic use of antioxidants is ensuring that topically administered agents remain in contact with the respiratory mucosa long enough to be effective. Normal mucociliary clearance removes topical medications from the respiratory tract within 15-20 minutes when it would be more desirable to maintain mucosal contact for hours to days. We propose to achieve targeted delivery to and retention of SOD and CAT (antioxidants) on respiratory epithelium using biodegradable polylactic acid (PLA) nanoparticles (NPs). We will simultaneously coat NPs with SOD and/or CAT and a targeting antibody (Ab) to mucin 1 (MUC1), which binds to the luminal surface of airway epithelial cells. Our preliminary studies demonstrate that HSNECs play a critical role in modulating the inflammatory response to environmental stimuli, and that targeting HSNECs with antioxidants can favorably impact the subsequent APC and T cell responses. Our central hypothesis is that smoke and DEP exposure experienced by OEF/OIF Veterans impacts HSNEC-APC communication via ROS and that this inflammatory response can be ameliorated through the use of antioxidant nanoparticles targeted at the respiratory epithelium. This hypothesis will be tested by the following specific aims: Specific Aim 1: To determine binding characteristics and epithelial toxicity of antibody-antioxidant- NPs. SOD and CAT will be covalently attached to PLA NPs simultaneously with antiMUC1 Ab. NPs with different ratios of SOD, CAT and antiMUC1 will be prepared and effects of the SOD:CAT:antiMUC1 ratios upon binding characteristics, function and toxicity will be evaluated in vitro using HSNEC cultures. Specific Aim 2: To determine in vitro efficacy of antioxidant NPs in inhibiting smoke and DEP induced inflammation. We will use optimized antiMUC1-SOD/CAT-NPs identified in Aim 1 to determine efficacy and ability to prevent smoke and DEP induced in vitro inflammatory response in HSNECs and APCs as the initial steps in the immune response. Specific Aim 3: To determine in vivo efficacy of antioxidant NPs on smoke and DEP exacerbated rhinosinusitis. We will use a murine model of allergic fungal rhinosinusitis with exacerbation by smoke or DEP exposure to determine the efficacy of antiMUC1-SOD/CAT-NPs. PUBLIC HEALTH RELEVANCE: NARRATIVE (RELEVANCE TO VETERANS HEALTH) Respiratory illnesses affect over two-thirds of veterans deployed to Iraq or Afghanistan and are among the most common medical conditions in Operation Enduring Freedom (OEF) and Operation Iraqi Freedom (OIF) veterans. Unique exposures to particulate matter in tobacco smoke and diesel exhaust particles from military vehicles likely contribute to this unusually high incidence and cause oxidative inflammation in the airway. Our strategic objective is to develop a topical medication, which could be used as a therapeutic measure against free radical pro-inflammatory action on respiratory epithelium that can persist even after removal of these environmental exposures. Our central hypothesis is that increased epithelial contact of nanoparticles conjugated with antibodies to the mucin 1 protein expressed on respiratory mucosa and with therapeutic antioxidants, superoxide and catalase, can prevent damage seen secondary to smoke and DEP exposures.